In vivo oxidative damage in rats is associated with barbiturate response but not other cytochrome P450 inducers
In vivo oxidative damage in rats is associated with barbiturate response but not other cytochrome P450 inducers
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DOI：
10.1124/mol.107.040238
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发表时间：
2007-12-01
影响因子：
3.6
通讯作者：
Guengerich, F. Peter
中科院分区：
文献类型：
--
作者：
Dostalek, Miroslav;Brooks, Joshua D.;Guengerich, F. Peter
先前发表的研究表明，细胞色素P450（P450）酶系统可以产生活性氧，并建议P450在氧化应激中的作用。然而，大多数研究都是在体外进行的，P450诱导和体内氧化损伤之间的潜在联系尚未用经验证的生物标志物进行严格探索。用典型的P450诱导剂预处理雄性Sprague-Dawley大鼠（β-萘醌、苯巴比妥（PB）、Aroclor 1254、异烟肼、双烯醇酮16 α-腈和氯贝特）或一般P450抑制剂1-氨基苯并三唑; P4501 A、-2B、-2E、-3A和-4A亚家族酶的诱导通过免疫印迹和使用光谱分析的1-氨基苯并三唑对P450的抑制来证实。PB和Aroclor 1254在体外显著增强丙二醛和H2 O2的生成和NADPH氧化，并在体内显著增强肝脏和血浆中的形成。其他一些治疗改变了体外参数，但没有改变体内参数。PB介导的肝脏和血浆F-2-异前列烷升高可被1-氨基苯并三唑消除，表明PB诱导的P450（s）参与F-2-异前列烷升高。体内氧化应激的标志物主要受PB和Aroclor 1254的影响，表明氧化损伤反应仅对巴比妥类诱导，可能与2B亚家族酶有关。这些研究定义了P450对体内氧化应激的贡献，因为这种现象相对有限，大多数P450基本上没有贡献。
Previously published studies have shown that cytochrome P450 (P450) enzyme systems can produce reactive oxygen species and suggest roles of P450s in oxidative stress. However, most of the studies have been done in vitro, and the potential link between P450 induction and in vivo oxidative damage has not been rigorously explored with validated biomarkers. Male Sprague-Dawley rats were pretreated with typical P450 inducers (beta-naphthoflavone, phenobarbital (PB), Aroclor 1254, isoniazid, pregnenolone 16 alpha-carbonitrile, and clofibrate) or the general P450 inhibitor 1-aminobenztriazole; induction of P4501A, -2B, -2E, -3A, and -4A subfamily enzymes was confirmed by immunoblotting and the suppression of P450 by 1-aminobenztriazole using spectral analysis. PB and Aroclor 1254 significantly enhanced malondialdehyde and H2O2 generation and NADPH oxidation in vitro and significantly enhanced formation in vivo, in both liver and plasma. Some of the other treatments changed in vitro parameters but none did in vivo. The PB-mediated increases in liver and plasma F-2-isoprostanes could be ablated by 1-aminobenztriazole, implicating the PB-induced P450(s) in the F-2-isoprostane elevation. The markers of in vivo oxidative stress were influenced mainly by PB and Aroclor 1254, indicative of an oxidative damage response only to barbiturate-type induction and probably related to 2B subfamily enzymes. These studies define the contribution of P450s to oxidative stress in vivo, in that the phenomenon is relatively restricted and most P450s do not contribute substantially.